Neochromosome, Inc., ConfidentialGene therapy aims to provide durable clinical benefit with a single treatment and has potential to cure a broadrange of disease conditions. Viral vector-based gene delivery accounts for the majority of current gene therapyclinical trials. Despite advancing the field, these vectors are limited by small DNA payloads (5-10kb), difficultvector production, tropism, immunogenicity and risk of carcinogenesis. Here we propose to develop a next-generation herpes simplex virus (HSV)-based vector system that overcomes these limitations. We aim toleverage HSV amplicon vectors, which can encode up to ~150kb of designer DNA sequence together with HSVpackaging and replication cis-signals. This simple design, which eliminates all viral protein-coding genes,ensures these vectors are non-toxic. Further, like other HSV vectors, amplicons remain extrachromosomal onceinside cells and pose no risk of insertional mutagenesis. While HSV amplicon gene therapy vectors have beenunder development for years, their use has been limited by safety concerns and manufacturing issues. Morespecifically, the absence of all viral genes necessitates the use of helper virus for amplicon packaging; to datean amplicon packaging system that completely excludes helper virus contamination has not been achieved. Herewe propose to build an amplicon packaging cell line for reliable production of 100% pure amplicon vectorpopulations devoid of helper gene sequences. We will demonstrate repeated passaging of amplicon vectors togenerate high titer stocks while maintaining 100% purity. Our vision is that large payload amplicon-mediatedgene delivery will enable the use of full-length human gene loci including introns and transcriptional regulatorysequences, allowing for natural spatial and temporal control of therapeutic gene expression. High capacityvectors that can accommodate large and multigene cassettes, such as HSV-1 derived systems, represent thefuture of gene therapy tools and will be essential to the treatment of complex genetic diseases particularly in thebrain.
Public Health Relevance Statement: Neochromosome, Inc., Confidential
PROJECT NARRATIVE
We will develop a packaging cell line for Herpes Simplex Virus (HSV)-based amplicon vectors that produces
high purity and titer viral particles carrying 150kb payloads for gene therapy. Such vectors will fill the need for
delivery of large genes and regulatory elements and/or gene clusters to treat a variety of diseases where such
tools are unavailable.
Project Terms: Back ; Dorsum ; Brain ; Brain Nervous System ; Encephalon ; Cell Line ; CellLine ; Strains Cell Lines ; cultured cell line ; Cell Nucleus ; Nucleus ; Cells ; Cell Body ; Clinical Trials ; Disease ; Disorder ; DNA ; Deoxyribonucleic Acid ; viral DNA ; virus DNA ; Engineering ; Episome ; Future ; Gene Activation ; Gene Cluster ; Gene Expression ; gene therapy ; DNA Therapy ; Gene Transfer Clinical ; Genetic Intervention ; gene-based therapy ; genetic therapy ; genomic therapy ; Genes ; Viral Genes ; Genome ; Helper Viruses ; Hippocampus (Brain) ; Ammon Horn ; Cornu Ammonis ; Hippocampus ; hippocampal ; Human ; Modern Man ; Infection ; Introns ; Intervening Sequences ; Mus ; Mice ; Mice Mammals ; Murine ; Neurons ; Nerve Cells ; Nerve Unit ; Neural Cell ; Neurocyte ; neuronal ; Production ; Risk ; Rodent ; Rodentia ; Rodents Mammals ; Safety ; Signal Transduction ; Cell Communication and Signaling ; Cell Signaling ; Intracellular Communication and Signaling ; Signal Transduction Systems ; Signaling ; biological signal transduction ; Technology ; Testing ; Genetic Transcription ; Gene Transcription ; RNA Expression ; Transcription ; Transfection ; Universities ; Viral Proteins ; Viral Gene Products ; Viral Gene Proteins ; virus protein ; Virion ; Virus Particle ; Virus Replication ; viral multiplication ; viral replication ; virus multiplication ; Virus ; Vision ; Sight ; visual function ; Insertional Mutagenesis ; Mediating ; promoter ; promotor ; DNA Sequence ; Custom ; base ; Site ; Clinical ; Phase ; Simplexvirus ; HSV ; Herpes Simplex Virus ; Herpes labialis Virus ; Herpesvirus 1 ; HSV-1 ; HSV1 ; Herpes Simplex Virus 1 ; Herpes Simplex Virus Type 1 ; herpes simplex i ; Ensure ; Evaluation ; Tropism ; Toxicity Tests ; Toxicity Testing ; Viral Packaging ; Virus Packagings ; Reporter ; tool ; Knowledge ; Complex ; cell type ; System ; Location ; Viral ; Cancer Induction ; carcinogenesis ; particle ; DNA Replication ; DNA Synthesis ; DNA biosynthesis ; professor ; Structure ; VP16 ; VP 16 ; Gene Expression Monitoring ; Gene Expression Pattern Analysis ; Transcript Expression Analyses ; Transcript Expression Analysis ; gene expression analysis ; gene expression assay ; transcriptional profiling ; Gene Expression Profiling ; Coding System ; Code ; Inflammatory Response ; protein expression ; Length ; immune competent ; Immunocompetent ; Non-dividing Cell ; Nondividing Cell ; Resting Cell ; Interphase Cell ; Mitotic ; Regulatory Element ; Gene Transduction Agent ; Gene Therapy Vectors ; Gene Transduction Vectors ; Gene Transfer ; Viral Vector ; Process ; Gene Delivery ; Development ; developmental ; vector ; immunogenicity ; design ; designing ; next generation ; HSV vector ; Herpes Simplex Virus Vector ; gene therapy clinical trial ; Population ; Impairment ; therapeutic gene ; gene therapeutics ; gene-based therapeutic ; gene-based therapeutics ; genes therapeutic ; genes therapeutics ; transgene expression ; transduction efficiency ; gene repair ; therapeutic DNA ; DNA-based therapeutics ; Genetic Diseases ; genetic condition ; genetic disorder ; Vectorial capacity ; Vectoral capacity ; genomic locus ; gene locus ; genetic locus ;
